Blackwell Science, LtdOxford, UK
ASJAnimal Science Journal1344-39412003 Blackwell Publishing Asia Pty Ltd
742April 2003
090
Influence of Starlink corn in cows
C. YONEMOCHIet al.
10.1046/j.1344-3941.2002.00090.x
Original Article8188BEES SGML

ORIGINAL ARTICLE
Influence of transgenic corn (CBH 351, named Starlink)on health condition of dairy cows and transfer of Cry9C
protein and cry9C gene to milk, blood, liver and muscle
Chisato YONEMOCHI,1 Takuo IKEDA,1 Chisato HARADA,1 Toyoko KUSAMA2 and Michito HANAZUMI1
1

Japan Scientific Feeds Association, Chiyoda-ku, Tokyo and 2National Fertilizer and Feed Inspection Station,
Saitama-shi, Japan

ABSTRACTThe influence of transgenic event CBH 351 (Starlink)-derived hybrid corn (SL) on the health condition, physiological
function and lactational performance of dairy cows as well as the transfer of the cry9C gene and Cry9C protein present
in SL to milk, blood, liver and muscles was examined, and compared with a diet containing non-transgenic (isogenic)
control corn (non-SL). After adapting to a diet containing non-SL for 2 weeks, four Holstein cows were assigned to eachof the non-SL and SL groups and were fed diets containing non-SL or SL, respectively, for 5 weeks. There were no
significant influences on the physiological condition, milk yield or serum biochemical and hematological values after
feeding with SL. There was also no influence on pH value, cell density of protozoa, or volatile fatty acid concentrationand composition of rumen fluids. In addition, no significant differences were observed on histopathological examinationof the major organs and tissues between the SL and non-SL groups. Moreover, the cry9C gene and Cry9C protein were
not detected by the polymerase chain reaction method and ELISA in the milk, blood, liver and muscles of the cows at
the end of the experiment.

KEYWORDS: lactating dairy cows, Starlink corn, transfer of cry9C and Cry9C.

INTRODUCTION

2002). Under these circumstances, it is inevitable that
GM corn will be used as feed in Japan. Therefore, after
confirming their safety, some GM corn varieties havebeen approved for use as feedstuffs by the Ministry ofAgriculture, Forestry and Fisheries of Japan.
The event CBH 351 corn, trade name Starlink (SL),is an insect-resistant corn which was developed in
France by AgrEvo (now Bayer CropScience, Kansas
City, MO, USA). It carries a novel gene from a naturally occurring soil bacterium, Bacillus thuringiensis ssp.
tolworthi, that encodes for a form of the Bt endotoxinknown as Cry9C protein (Schnepf et al. 1998).

Attempts to make genetically modified organisms(GMO) were started in the 1970s and a ‘delayed softening tomato’ was approved by the United States Foodand Drug Administration (USFDA) in 1991. At thepresent time, 53 varieties of GMO crops and vegetables
including corn, rapeseed, cotton, soybean, beet,
potato, squash and chicory have been developed(USFDA 2002). Among them, 16 varieties of genetically modified corn (GM corn) that carry resistance tochemical herbicides and insects have been approvedby the USFDA and the planted area of GM cornincreased to 34% of the total planted area in the USA
in the year 2002 (USDA 2002).
In Japan, more than 70% of feedstuffs are imported
from foreign countries. In particular, more than 95%
of imported feed corn comes from the USA (MAFF

Animal Science Journal (2003) 74, 81–88

Correspondence: Chisato Yonemochi, Kanda-Surugadai
1–2, Chiyoda-ku, Tokyo 101–0062, Japan. (Email:yone@kashikyo.or.jp)
Received 4 June 2002; accepted for publication 11 October
2002.

81

C. YONEMOCHI et al.

Because Cry9C protein is not digested completely invitro and can be an allergen for humans, it has been
approved only for use as a feedstuff and for alcohol
production in the USA by the USFDA.
In contrast, SL is not approved for either food or
feedstuff use in Japan and the discovery of SL in foodand feedstuffs in Japan caused social anxiety relating
to the safety of SL (MAFF 2000).
Bayer CropScience presented various data concerning the safety of SL to the USFDA. However, these data
did not contain any results of feeding tests or the
possibility of transferring the cry9C gene and Cry9C
protein to animal tissues (USEPA 1998). Therefore, it
is very important to confirm the effects of SL on thehealth of animals and the possibility of transferring thecry9C gene and Cry9C protein to animal products. The
authors have already confirmed that SL does not affectthe health of broiler chicks and that the cry9C gene and
Cry9C protein is not transferred to the meat, liver orblood of broilers given free access to a diet containing
70% SL for 7 weeks (Yonemochi et al. 2002).
Following on from this work, the present study was
conducted to confirm the effect of SL on the performance and physiological function of lactating dairy
cows and the possibility of transferring the cry9C gene
and Cry9C protein derived from SL to the milk, blood,
liver and muscles of dairy cows fed a diet with 35%
SL for 5 weeks.

Before formulating the experimental diets, both SL
and non-SL were ground with a hummer mill througha 1 mm screen.

Diet
The compositions of the experimental diets are shownin Table 2. Each experimental diet contained 35% of
either non-SL or SL. The cows were fed the experimental diet twice a day (at 09.00 hours and17.00 hours), to provide 110% of the total digestible
nutrients requirement for maintenance and the production of milk according to the Japanese FeedingStandard for Dairy Cattle (MAFF 1999) (Table 3).

Experimental design
The experiment was performed between November 3
and December 22, 2000 in our Research Center. The
characteristics of the dairy cows are shown in Table 3.
During the 2-week pre-experimental period, eight
Holstein cows were supplied the non-SL diet to adapt
to the experimental environment, and then they wereallotted to two groups, non-SL and SL dietary groups,
each containing four cows. Each cow was tethered bya neck chain in a stall furnished with a 1.3 ¥ 1.8 m
rubber mat. An empty stall was left between stalls toprevent cross contamination. The SL group was separated by a vinyl sheet curtain from the non-SL group.
Tap water was given ad libitum.

MATERIALS AND METHODS

Table 2 Ingredients and chemical compositions of
the experimental dietCorn grain(%)

The two corn varieties, non-SL and SL, used in thepresent study were supplied by Bayer CropScience. In
the non-SL, neither the cry9C gene nor the Cry9C
protein were detected by the polymerase chain reaction method (PCR) or ELISA, respectively. The cry9C
gene was carried by 89.3% of the SL grains.
There were no differences in chemical composition
between non-SL and SL, as shown in Table 1.

Ingredients
Corn (SL or non-SL)
Soybean meal
Wheat bran
Soybean hull
Calcium carbonate
Dicalcium phosphate
Sodium chloride
Trace mineral premix†
Vitamin ADE premix‡
Timothy hay
Alfalfa hay cube
Calculated analysis§
Neutral detergent fiber
Digestible crude protein
Total digestible nutrients

Table 1 Proximate analysis of SL and non-SL corn (%)

Moisture
Crude protein
Ether extract
Nitrogen free extract
Crude fiber
Crude ashSL

Non-SL

10.6
7.7
4.4
74.3
1.6
1.4

10.4
7.8
4.3
74.6
1.5
1.4

31.5
12.8
66.1

†Contained (g/kg of premix): manganese 50, iron 50, copper 10,
zinc 60, iodine 1. ‡Contained (IU or mg/kg of premix): vitamin A
10 000 IU, vitamin D3 2000 IU, DL-a-tocopherol acetate 10 mg.
§Based on MAFF 1995. SL, Event CBH 351 corn (trade nameStarlink); Non-SL, Non-transgenic (isogenic) control corn.

SL, Event CBH 351 corn (trade name Starlink); Non-SL,
Non-transgenic (isogenic) control corn.

Animal Science Journal (2003) 74, 81–88

35.0
8.75
2.7
2.5
0.5
0.25
0.2
0.05
0.05
35.0
15.0

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Influence of Starlink corn in cows

Table 3 Characteristics of the cows and feed intake

Group

Cow

Age

No. ofparturitions

Stage oflactation(days after
parturition)

Body weight
(kg)
Start

End

Feed intake (kg/day)
Concentrate

Timothyhay

Alfalfa hay
cube

non-SL
A
B
C
D

4
4
5
3

2
2
3
1

151
27
298
209

536
615
608
689

544
616
615
696

11.7
9.1
8.6
11.8

8.2
6.3
6.0
8.3

3.5
2.7
2.6
3.5

E
F
G
H

7
3
6
2

4
1
4
1

62
293
255
169

766
546
700
627

789
567
702
647

10.7
10.2
8.8
10.0

7.5
7.1
6.1
7.0

3.2
3.0
2.6
3.0

SL

SL, Event CBH 351 corn (trade name Starlink); Non-SL, Non-transgenic (isogenic) control corn.

Health condition

Histopathological examination: At the end of theexperiment, the cows were humanely slaughteredafter anesthetizing with Rompun 2% solution (Bayer
AG, Leverkusen, Germany) and then bled through thejugular vein. Tissue samples from the cerebrum, cerebellum, lung, heart, liver, kidney, pancreas, rumen,
duodenum and jejunoileum were collected. After the
specimens were fixed, thin sections were cut, stainedwith hemotoxylin and eosin, and examined under alight microscope.

The body weight of each cow was measured at the
beginning and end of the experiment. The health status was diagnosed by observing the appearance of theanimal’s body and feces at each milking time.

Milk production
The cows were milked at 09.00 hours and 17.00 hours
by a bucket milker. The milk yield per day was calculated from the total amount of milk produced during
one week or the entire experimental period.

Analysis of rumen fluid
The rumen fluid was collected orally through a stomach tube at 13.00 hours at the beginning and end of
the experiment. Its composition, pH value, protozoa
counts and total volatile fatty acid (VFA) concentration were examined. The VFA were analyzed by gas
chromatography, using a glass column packed with
Chromosorb W (80/100 mesh) coated with 5%
Thermo 1000 and 0.5% H3PO4 (Wako Pure Chemical
Industries, Tokyo, Japan). The protozoa were counted
according to the method of Imai and Katsuno (1977).

Serum biochemical, hematological andhistopathological examination
Serum biochemical examination: Blood was collectedfrom the jugular vein at the beginning and end of theexperiment and the sera were separated. Lactate dehydrogenase (LDH), glutamic-oxalacetic transaminase(GOT), glutamic-pyruvic transaminase (GPT), alkaline
phosphatase (ALP), blood urea nitrogen (BUN), neutral lipid (NL), total bilirubin (T-Bili), total cholesterol(T-Chol), total protein (T-P), albumin (Alb) and glucose (Glu) were determined by an autoanalyzer(model JCB MB8, Nihon Denshi, Tokyo, Japan). Globulin (Glob) and the Alb/Glob (A/G) ratio were calculated on the basis of T-P and Alb concentrations.
Hematological examination: Blood samples werecollected from the jugular vein. Hemoglobin (Hb;azido-methemoglobin method), hematocrit (Ht; capillary method), red blood cells (RBC; Coulter Counter,model ZF; Coulter Electronics, Miami, FLO, USA), andwhite blood cells (WBC; indirect method) were
determined.

Animal Science Journal (2003) 74, 81–88

Detection of cry9C gene and Cry9C protein
in blood and tissue
At the end of the experiment, the blood, muscles andliver were collected individually.
Detection of the cry9C gene by PCR: Deoxyribonucleic acid (DNA) was isolated from the blood, liver andmeat according to the procedures described in ourprevious report (Yonemochi et al. 2002). Deoxyribonucleic acid (DNA) was isolated from milk according
to the procedures of Lipkin et al. (1993), except thatthe milk somatic cells were lysed by incubation in83

C. YONEMOCHI et al.

RESULTSboiling water for 10 min. Gel electrophoresis, PCR
amplification, and the detection of PCR products ongels were performed by the methods described in ourprevious report (Yonemochi et al. 2002).
Detection of Cry9C protein by ELISA: The blood,
liver and meat were treated according to the procedures described in our previous report (Yonemochi
et al. 2002). Milk was treated in the same way as
blood. The determination of Cry9C protein in the samples was as described in our previous report (Yonemochi et al. 2002).

Health condition and milk production
There were no refusals of feed by any of the cows
during the experimental period and no abnormalities
in the health of the cows were observed.
Changes in milk yield during the experimental
period are presented in Table 4. No significant differences in milk yield were found between the SL andnon-SL groups during any week.

Serum biochemical values
Statistical analysis

The serum biochemical values are presented inTable 5. There were no significant differences in LDH,
GOT, ALP, T-Chol, T-P and the A/G ratio between the
non-SL and SL groups and between the first and lastdays of the experiment. Also there were no differencesin Glu and Alb between non-SL and SL groups. Glucose was higher and Alb was lower on the first day
than on the last day. In contrast, GPT and BUN of theSL group were lower than those of the non-SL group,
and GPT on the first day was higher than on the lastday. However, these variations ranked within thenormal values for a lactating dairy cow (Takahashi &Data on milk yield were analyzed by a one-way
analysis of variance (Yoshida 1983). Data on serum
biological values, hematological values and rumen
fermentation parameters were analyzed by a two-way
analysis of variance. Then they were presented in a
way in which the main effect of the group (SL groupvs non-SL group) and sampling periods (the first dayof the experiment vs the last day of the experiment)
were shown, and the group by sampling periods interaction was included in the model. Significant differences were tested at a level of P < 0.05.
Table 4 Change of milk yields (kg/head/day)Group

non-SLSL

Week

Average

1st

2nd

3rd

4th

5th

25.8 ± 6.4†
25.0 ± 3.3

25.0 ± 2.7
28.0 ± 6.0

24.6 ± 3.1
27.2 ± 5.9

23.9 ± 3.0
27.0 ± 5.4

23.4 ± 2.5
26.0 ± 4.0

24.5 ± 3.4
26.6 ± 4.4†Values are means ± SD (n = 4). SL, Event CBH 351 corn (trade name Starlink); Non-SL, Non-transgenic (isogenic) control corn.

Table 5 Serum biochemical values for cows at the start and end of the experiment

Sampling periods
GroupsFirst day of experiment

Last day of experiment

Probabilities‡

non-SLSL

non-SLSL

Group

Sampling periods

Interaction

LDH (IU/L)
GOT (IU/L)GPT (IU/L)
BUN (mg/dL)
NL (mg/dL)
T-Bili (mg/dL)
ALP (IU/L)
T-Chol (g/dL)
Glu (mg/dL)
T-P (g/dL)
Alb (g/dL)
A/G

1633 ± 150†
60.3 ± 9.5
22.5 ± 3.3
17.3 ± 2.8
<25
<0.30
186 ± 199
159 ± 30
82.5 ± 10.9
7.5 ± 0.7
3.7 ± 0.4
1.05 ± 0.28

1643 ± 289
51.0 ± 10.4
15.0 ± 3.9
13.5 ± 3.8
<25
<0.30
96 ± 43
164 ± 33
79.8 ± 5.0
7.5 ± 0.5
3.8 ± 0.2
1.04 ± 0.16

1752 ± 411
57.8 ± 10.2
16.0 ± 2.7
15.0 ± 1.2
<25
<0.30
184 ± 86141 ± 33
69.8 ± 4.9
7.5 ± 0.4
4.0 ± 0.2
1.22 ± 0.41

1712 ± 223
45.8 ± 11.9
13.0 ± 3.8
11.8 ± 1.3
<25
<0.30
86 ± 27150 ± 33
76.5 ± 3.9
7.5 ± 0.4
4.0 ± 0.1
1.19 ± 0.18

NS
NS
*
*
–§
–
NS
NS
NS
NS
NS
NS

NS
NS
*
NS
–
–
NS
NS
*
NS
*
NS

NS
NS
NS
NS
–
–
NS
NS
NS
NS
NS
NS

†Values are mean ± SD (n = 4). ‡NS, Not significant (P > 0.05); *, Significant (P £ 0.05); §not applicable. SL, Event CBH 351 corn (trade nameStarlink); Non-SL, Non-transgenic (isogenic) control corn. LDH, lactate dehydrogenase; GOT, glutamic-oxalacetic transaminase; GPT, glutamicpyruvic transaminase; BUN, blood urea nitrogen; NL, neutral lipid; T-Bili, total bilirubin; ALP, alkaline phosphatase; T-Chol, total cholesterol;Glu, glucose; T-P, total protein; Alb, albumin; A/G, albumin/globulin.

Animal Science Journal (2003) 74, 81–88

84

Influence of Starlink corn in cows

Table 6 Hematological values for cows at the start and end of the experiment

Sampling periods
Groups6

RBC (10 /mL)
WBC (103/mL)
Hb (g/dL)
Ht (%)First day of experiment

Last day of experiment

Probabilities‡

non-SLSL

non-SLSL

Group

Sampling periods

Interaction

7.01 ± 0.39†
9.6 ± 1.6
10.2 ± 1.1
32.9 ± 1.7

7.36 ± 1.16
8.4 ± 1.1
10.8 ± 1.1
34.8 ± 2.2

8.33 ± 0.46
7.9 ± 1.8
10.1 ± 0.7
35.9 ± 1.6

8.05 ± 0.83
6.8 ± 0.6
10.0 ± 0.5
37.5 ± 2.5

NS
NS
NS
NS

*
*
NS
*

NS
NS
NS
NS

†Values are mean ± SD (n = 4). ‡NS, Not significant (P > 0.05); *, Significant (P £ 0.05). RBC, red blood cells; WBC, white blood cells;
Hb, hemoglobin; Ht, hematocrit.

Table 7 Characteristics of rumen fluid from cows at the start and end of the experiment

Sampling periods
GroupsFirst day of experiment

Last day of experiment

Probabilities‡

non-SLSL

non-SLSL

Group

Sampling
periods

Interaction

pH
Protozoa count (105/mL)
Total VFA (mmol/L)
VFA composition (mol%)
Acetic acid
Propionic acid
Butyric acid
Others
A/G

7.1 ± 0.2†
2.98 ± 1.60
209.9 ± 26.4

7.2 ± 0.1
3.63 ± 1.38
208.1 ± 20.5

7.2 ± 0.3
1.94 ± 0.29
200.6 ± 58.6

7.3 ± 0.3
3.02 ± 1.26
202.1 ± 51.7

NS
NS
NS

NS
NS
NS

NS
NS
NS

57.4 ± 0.8
18.3 ± 1.6
16.4 ± 0.6
7.9 ± 1.1
3.15 ± 0.31

56.9 ± 2.2
18.0 ± 2.0
17.9 ± 2.3
7.4 ± 0.6
3.21 ± 0.47

55.8 ± 1.2
20.1 ± 1.7
15.9 ± 1.1
8.3 ± 1.4
2.80 ± 0.28

55.8 ± 0.9
18.1 ± 2.1
17.9 ± 1.9
8.2 ± 0.6
3.12 ± 0.41

NS
NS
NS
NS
NS

NS
NS
NS
NS
NS

NS
NS
NS
NS
NS

†Values are mean ± SD (n = 4). ‡NS, Not significant (P > 0.05). VFA, volatile fatty acids; A/G, albumin/globulin.

Itagaki 1975; Nakamura 1976; Sonota 1978), andthere were no significant differences in the interaction
between the groups and the sampling periods.

in the pH value, protozoa count, total VFA concentration and its composition between the non-SL and SL
groups on the first and last days of the experiment.
The interaction relationship between the groups andsampling periods were not significant.

Hematological values
The hematological values for both groups are presented in Table 6. There were no significant differencesin Hb between the non-SL and SL groups on the firstor last days of the experiment. Although there wereno significant differences in RBC, WBC and Ht
between non-SL and SL groups, RBC and Ht on thefirst day were significantly lower than on the last day.
White blood cells on the first day was significantly
higher than on the last day. These variations were also,
however, within the normal values for a lactating
dairy cow (Takahashi & Itagaki 1975; Nakamura 1976;
Sonota 1978), and there were no significant differences in interaction between the groups and the sampling periods.

Necropsy finding and histopathologicalobservation
Necropsy revealed several abnormalities. In the nonSL group, a liver abscess was found in Cow C. In the
SL group, a lipoma around the colon was found in
Cow E, a simple polyp on the rumen and hydronephrosis were found in Cow G, and a simple polyp was
found on the rumen of Cow H. No specific abnormality was observed in any of the cows.
The results of the histopathological examination arepresented in Table 8. No significant lesions were foundin the cerebrum, cerebellum, heart, pancreas, duodenum or jejunoileum of any of the cows. Many of thelesions described in Table 8 are often found in specimens from conventional cows. Furthermore, theselesions were all fairly minor and not considered tohave any effect on their physiological function.

Characteristics of rumen fluid
The results of the rumen fluid examination are presented in Table 7. There were no significant differences

Animal Science Journal (2003) 74, 81–88

85

C. YONEMOCHI et al.

Table 8 Results of histopathological observation†

Organ

non-SL
Cow

SL
Findings

Lung
Liver

C

D
KidneyC
D

Left lobe: bleeding in lobe (mild). Scatteredleukocyte infiltration inside Glisson’s
capsule (middle degree)
Right lobe: scattered leukocyte infitration
inside lobe and Glisson’s capsule (minor).
Left kidney: infiltration of lymph-like cell in
cortex (mild).
Left kidney: fattening of Bowman’s capsule,minor infiltration of lymph-like cell insurrounding (minor). Cortex: nest-like
infiltration of lymph-like cell which acrossvertical linear, necrosis of uriniferous
tubule (middle degree).

Rumen

Cow

Findings

G

Deposit of hemosiderin on alveolus wall (migrationof leukocyte on the part of bronchiole, minor).
Right lobe: scattered lymph-like cell infiltration
between lobes (minor).

H

E
G

G, H

Right kidney: scattered minor fattening ofBowman’s capsule (mild).
Left kidney: infitration of lymph-like cell in cortex,
fattening of Bowman’s capusule (minor). Depositof hemosiderin in urinary tubular cell (mild).
Expansion of part of collecting tubule (minor).

Fibroepithelioma (simple polyp) from mucous
exists.

†Only the regions and cows that showed abnormalities are presented. SL, Event CBH 351 corn (trade name Starlink); Non-SL, Non-transgenic(isogenic) control corn.

Probability of transfer of cry9C gene and
Cry9C protein to blood, liver, muscle
and milk

examination, because there were also no significant
differences in the interaction between groups andsampling periods. The observed total VFA concentration was relatively high in both groups compared withthe average, but these observed values ranked within
normal values (Akay & Jackson 2001).
Although there has been no report on the effect ofSL on the lactating performance of dairy cows, Folmer
et al. (2000) and Donkin et al. (2000) reported theeffect of other GM corn varieties (Bt-11, Folmer et al.
2000, and Roundup Ready, Donkin et al. 2000) on thelactating performance of dairy cows. These studies
were conducted to elucidate the milk yield and composition using a Latin squire design (Folmer et al.
2000) or double reverse feeding (Donkin et al. 2000)
which are commonly used for feeding experimentswith dairy cows.
In contrast, the present study was conducted to elucidate whether or not the cry9c gene and Cry9C protein were transported into milk, blood, liver andmuscle. In the present study, parallel comparative
feeding was employed instead of reverse feeding,because transfer of the cry9C gene and Cry9C protein
into the tissues had to be determined at the end ofexperiment.
There were no differences in milk yield between the
SL and non-SL groups in the present study. This result
was in agreement with those of Folmer et al. (2000)
and Donkin et al. (2000). Moreover, in the present

On the final day of the experiment, neither the cry9C
gene, nor the Cry9C protein, were detected in themilk, blood, liver or muscles of the cows in either the
non-SL or SL groups, by PCR and ELISA, respectively(data not shown).
In all cases, DNA strands of 315-bp specific for thebovine growth hormone region were amplified by
PCR from DNA obtained from the milk, blood, liver
and muscles in both groups, while DNA strands of
379-bp specific for the cry9C region were not amplified. The agarose gel electrophoresis patterns of PCR
products amplified from milk DNA are shown in
Fig. 1. The same results were obtained for samples
from the blood, liver and muscles (data not shown).
These results indicate that the cry9C gene and Cry9C
protein contained in SL corn do not transfer to themilk, blood, liver and muscles.

DISCUSSION
In the present study, the health condition of dairy
cows was not affected when fed for 5 weeks on a dietwith 35% SL corn. Furthermore, there were no differences in serum biochemical and hematological values, and feeding SL did not affect histpathological

Animal Science Journal (2003) 74, 81–88

86Influence of Starlink corn in cows(a)Fig. 1 Detection of growth
hormone gene and cry9C gene inmilk of dairy cows by PCR. PCR
product (315 bp) specific for growth
hormone gene (A), and PCR product(379 bp) specific for cry9C generegion in SL (B). Two samples ofmilk for each cow were loaded onto
the gel. M, 100 bp Ladder; Lanes
1–8, milk of cows from the non-SLgroup; lanes 9–16, milk of cows
from the SL group; N, negative
control (no template); P, positive
control (PCR product amplified
from Starlink corn).

(b)

study, the cry9c gene and Cry9C protein were notdetected by PCR and ELISA in milk, blood, liver andmuscle.
In conclusion, the results obtained in the present
study indicate that feeding SL does not influence thehealth condition or physiological function in lactating
dairy cows, compared with those fed non-SL. The cry9cgene and Cry9C are not transferred to milk, blood,
liver and muscle of dairy cows.

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ACKNOWLEDGMENT
This work was funded by a grant from the Agriculture
Livestock Industry Corporation.

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